title: A whole-organ regenerative medicine approach for liver replacement.
abstract: OBJECTIVE
  The therapy of choice for end-stage liver disease is whole-organ liver transplantation, but this option is limited by a shortage of donor organs. Cell-based therapies and hepatic tissue engineering have been considered as alternatives to liver transplantation, but neither has proven effective to date. A regenerative medicine approach for liver replacement has recently been described that includes the use of a three-dimensional organ scaffold prepared by decellularization of xenogeneic liver. The present study investigates a new, minimally disruptive method for whole-organ liver decellularization and three different cell reseeding strategies to engineer functional liver tissue.
  
  
  METHODS
  A combination of enzymatic, detergent, and mechanical methods are used to remove all cells from isolated rat livers. Whole-organ perfusion is used in a customized organ chamber and the decellularized livers are examined by morphologic, biochemical, and immunolabeling techniques for preservation of the native matrix architecture and composition. Three different methods for hepatocyte seeding of the resultant three-dimensional liver scaffolds are evaluated to maximize cell survival and function: (1) direct parenchymal injection, (2) multistep infusion, or (3) continuous perfusion.
  
  
  RESULTS
  The decellularization process preserves the three-dimensional macrostructure, the ultrastructure, the composition of the extracellular matrix components, the native microvascular network of the liver, and the bile drainage system, and up to 50% of growth factor content. The three-dimensional liver matrix reseeded with the multistep infusion of hepatocytes generated ∼90% of cell engraftment and supported liver-specific functional capacities of the engrafted cells, including albumin production, urea metabolism, and cytochrome P450 induction.
  
  
  CONCLUSIONS
  Whole-organ liver decellularization is possible with maintenance of structure and composition suitable to support functional hepatocytes.
authors.affiliation: Transplantation Section, Department of Surgery, Children's Hospital of Pittsburgh, Center for Innovative Regenerative Therapies, University of Pittsburgh, Pennsylvania, USA. sotogutierreza@upmc.edu
authors.forename: Alejandro
authors.initials: A
authors.lastname: Soto-Gutierrez
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Chris
authors.initials: C
authors.lastname: Medberry
authors.forename: Ken
authors.initials: K
authors.lastname: Fukumitsu
authors.forename: Denver
authors.initials: D
authors.lastname: Faulk
authors.forename: Hongbin
authors.initials: H
authors.lastname: Jiang
authors.forename: Janet
authors.initials: J
authors.lastname: Reing
authors.forename: Roberto
authors.initials: R
authors.lastname: Gramignoli
authors.forename: Junji
authors.initials: J
authors.lastname: Komori
authors.forename: Mark
authors.initials: M
authors.lastname: Ross
authors.forename: Masaki
authors.initials: M
authors.lastname: Nagaya
authors.forename: Eric
authors.initials: E
authors.lastname: Lagasse
authors.forename: Donna
authors.initials: D
authors.lastname: Stolz
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Strom
authors.forename: Ira J
authors.initials: IJ
authors.lastname: Fox
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Badylak
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology / Q000637:transplantation*; D019164:Liver, Artificial; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques / Q000379:methods*; D011474:Prosthesis Design; D051381:Rats; D017207:Rats, Sprague-Dawley; D054457:Tissue Scaffolds